通过在吸水链霉菌中过表达调控基因来提高阿霉素的产量。
Improvement in doxorubicin productivity by overexpression of regulatory genes in Streptomyces peucetius.
机构信息
Institute of Biomolecule Reconstruction, Department of Pharmaceutical Engineering, Sun Moon University, #100, Kalsan-ri, Tangjeong-myeon, Asansi, Chungnam 336-708, Republic of Korea.
出版信息
Res Microbiol. 2010 Mar;161(2):109-17. doi: 10.1016/j.resmic.2009.12.003. Epub 2010 Jan 4.
Biosynthesis of doxorubicin (DXR) is tightly regulated, limiting its production in Streptomyces peucetius cultures. The regulatory genes dnrN, dnrI, afsR, and metK1-sp from S. peucetius ATCC 27952 were cloned into the pIBR25 expression vector under the control of the strong ermE* promoter to enhance DXR production. The constructed expression plasmids, pNI25 (with dnrN-dnrI), pNIS25 (with dnrN-dnrI-metK1-sp), pNIR25 (with dnrN-dnrI-afsR), pRS25 (with afsR-metK1-sp) and pNIRS25 (with dnrN-dnrI-afsR-metK1-sp), were transformed into S. peucetius. The recombinant strains NI, NIS and NIR produced greater amounts of DXR than the parental strain with an increment of 1.2-fold by pNI25, 1.4-fold by pNIS25 and 4.3-fold by pNIR25, whereas pRS25 and pNIRS25 had no significant effect on DXR production. We also studied the transcriptional level of overexpressed regulatory genes and relative production of DXR, daunorubicin (DNR) and epsilon-rhodomycinone (RHO) in each recombinant strain.
阿霉素（DXR）的生物合成受到严格调控，限制了其在变铅青链霉菌培养物中的产量。变铅青链霉菌 ATCC 27952 的调节基因 dnrN、dnrI、afsR 和 metK1-sp 被克隆到 pIBR25 表达载体中，受强 ermE*启动子的控制，以提高 DXR 的产量。构建的表达质粒 pNI25（带有 dnrN-dnrI）、pNIS25（带有 dnrN-dnrI-metK1-sp）、pNIR25（带有 dnrN-dnrI-afsR）、pRS25（带有 afsR-metK1-sp）和 pNIRS25（带有 dnrN-dnrI-afsR-metK1-sp）被转化到变铅青链霉菌中。重组菌株 NI、NIS 和 NIR 产生的 DXR 量比原始菌株多，pNI25 增加了 1.2 倍，pNIS25 增加了 1.4 倍，pNIR25 增加了 4.3 倍，而 pRS25 和 pNIRS25 对 DXR 产量没有显著影响。我们还研究了每个重组菌株中过表达调节基因的转录水平和 DXR、柔红霉素（DNR）和 ε-罗红霉素（RHO）的相对产量。
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